serum, plasma, tissue homogenates, cell lysates, cerebrospinal fluid, cell culture supernates and other biological fluids
Assay Length:
2h
Research Area:
Signal transduction;Endocrinology;Neuro science;Hormone metabolism;
Test principle:
This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Rat bEP protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat bEP. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Rat bEP in the samples is then determined by comparing the OD of the samples to the standard curve.
標(biāo)準(zhǔn)曲線
Concentration (pg/mL)
OD
Corrected OD
1000.00
0.183
500.00
0.275
250.00
0.446
125.00
0.846
62.50
1.389
31.25
1.201
15.63
1.624
0.00
2.236
精密度
Intra-assay Precision (Precision within an assay):CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays):CV%<10%
Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
回收率
Matrices listed below were spiked with certain level of recombinant bEP and the recovery rates were calculated by comparing the measured value to the expected amount of bEP in samples.
Matrix
Recovery range
Average
serum(n=5)
87-99%
93%
EDTA plasma(n=5)
82-97%
90%
Heparin plasma(n=5)
88-106%
97%
線性
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of bEP and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.